Abstract:The cuttings from 5yearold and 30yearold chestnut trees were firstly used to compare their rooting, and then cuttings from 30yearold chestnut trees were studied in May, June and July, including the effects of different IBA concentrations (0, 200 and 500 mg/L). During the cutting process, the rooting of various treatments, the carbon and nitrogen metabolites, browning related substances and anatomical structures were investigated before cutting (initial stems) and after cutting (calluses), and then the rooting ability of cuttings in different mother trees and different months were discussed, which would lay a foundation for the study of rooting of chestnut cuttings. The results showed: (1) the starch and soluble sugars in the initial stems and calluses of the 5yearold chestnut were significantly higher than those of the 30yearold chestnut, while the soluble proteins were contrary, but GABA did not change significantly between different mother trees. For 30yearold chestnut cuttings, the contents of soluble proteins, GABA and soluble sugars decreased in the initial stems, but GABA, soluble sugars and starch increased gradually in the calluses with the increasing months. (2) The contents of tannins, phenols and flavonoids in the initial stems and calluses of chestnut cuttings were almost less in 30yearold trees than those of 5yearold trees. Except for the significant increase in flavonoids in the calluses in July, there were no significant changes in the contents of tannins, phenols and flavonoids in different months. (3) The observation of rooting status and anatomy indicated that the callus rate and rooting rate in 5yearold chestnut cuttings were higher than those of the 30yearold ones, and the callus rate was the highest in June. Chestnut cuttings belonged to the type of induced rooting, and the annular thickwalled tissue existed in the cortex. The lignin content of the 5yearold chestnut cuttings was lower than that of the 30yearold ones, but the number of cambium layers was contrary. Our study indicated that chestnut cuttings were severely browned, resulting in a lower rooting rate; there were significant differences in carbon and nitrogen metabolism related compounds in the cuttings from the different mother trees and different months; the browningrelated substances had significant differences in the cuttings from differentage mother trees, but there was little change between different months.